Ether derivatives of the FK-506 related macrolide L-683,742 (C31-0-desmethyl ascomycin) have been prepared by alkylation with various alkyl2,2,2-trichlomacetimidates. The in vitro immunosuppressive activity of these analogs is dependent both on the size and site of attachment (C31-0 vs. C32-0) of the ether group.